http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&feed=atom&action=historyTeam:NYMU-Taipei/project/1c - Revision history2024-03-29T15:31:36ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=401400&oldid=prevTree1220 at 03:57, 18 October 20142014-10-18T03:57:00Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>(OD600=2.647)(2.0 rcf, 7min) </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>(OD600=2.647)(2.0 rcf, 7min) </li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>J23100+yebF+endolysin+B0015:overnight culture supernatant </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>J23100+yebF+endolysin+B0015:overnight culture supernatant </li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li><del class="diffchange diffchange-inline">J23100+yebF+endolysin+B0015 </del>(OD600=2.900)(2.0 rcf, 7min)</li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li>(OD600=2.900)(2.0 rcf, 7min)</li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Western blot filter paper:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Western blot filter paper:</p></div></td></tr>
</table>Tree1220http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=401008&oldid=prevTree1220 at 03:54, 18 October 20142014-10-18T03:54:05Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ampicillin : 100ng/ul</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ampicillin : 100ng/ul</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>BHI broth</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>BHI broth</li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li><del class="diffchange diffchange-inline">J23100</del>+<del class="diffchange diffchange-inline">yebF</del>+<del class="diffchange diffchange-inline">B0015</del>: J23100+yebF+B0015 overnight culture supernatant</li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li><ins class="diffchange diffchange-inline">J</ins>+<ins class="diffchange diffchange-inline">Y</ins>+<ins class="diffchange diffchange-inline">B</ins>: J23100+yebF+B0015 overnight culture supernatant</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>(OD600=2.647)(2.0 rcf, 7min) </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>(OD600=2.647)(2.0 rcf, 7min) </li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>J23100+yebF+endolysin+B0015:overnight culture supernatant </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>J23100+yebF+endolysin+B0015:overnight culture supernatant </li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Western blot filter paper:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Western blot filter paper:</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li>Positive control: Ampicillin (100ng/ul),<del class="diffchange diffchange-inline">50ml</del></li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li>Positive control: Ampicillin (100ng/ul),<ins class="diffchange diffchange-inline">50ul</ins></li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li>Negative control: BHI, 50ml /J23100+yebF+B0015, <del class="diffchange diffchange-inline">50ml</del></li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li>Negative control: BHI, 50ml /J23100+yebF+B0015, <ins class="diffchange diffchange-inline">50ul</ins></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Testing:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Testing:</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li>J23100+yebF+endolysin+B0015, <del class="diffchange diffchange-inline">50ml</del></li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li>J23100+yebF+endolysin+B0015, <ins class="diffchange diffchange-inline">50ul</ins></li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li>J23100+yebF+endolysin+B0015, <del class="diffchange diffchange-inline">100ml</del></li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li>J23100+yebF+endolysin+B0015, <ins class="diffchange diffchange-inline">100ul</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li>J23100+yebF+endolysin+B0015, 150ul</ins></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ul></div></td></tr>
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</table>Tree1220http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=392471&oldid=prevTree1220 at 02:55, 18 October 20142014-10-18T02:55:05Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Before we check the functionality of the entire construct, we need to test the YebF protein for its ability to transport attached parts out of the membrane. We began by constructing two circuits:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Before we check the functionality of the entire construct, we need to test the YebF protein for its ability to transport attached parts out of the membrane. We began by constructing two circuits:</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/d/d3/NYMU14_target_YebF_t.png' style='width: 550px;'></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/d/d3/NYMU14_target_YebF_t.png' style='width: 550px;'></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>These circuits are then transformed into <i>E. coli</i>, cultivated in liquid culture, and centrifuged. Afterwards, the supernatants are analyzed for flourescence intensity and their OD value. If YebF works as we expected, we should see the first circuit successfully secrete RFP into the supernatant :</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>These circuits are then transformed into <i>E. coli</i>, cultivated in liquid culture, and centrifuged. Afterwards, the supernatants are analyzed for flourescence intensity <ins class="diffchange diffchange-inline">fluorescence intrnsity </ins>and their OD value. If YebF works as we expected, we should see the first circuit successfully secrete RFP into the supernatant :</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/a/a6/NYMU14_target_YebF_rfp.png' style='width: 220px;margin-left: 40px;'></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/a/a6/NYMU14_target_YebF_rfp.png' style='width: 220px;margin-left: 40px;'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>While the control should only contain RFP which is not exported:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>While the control should only contain RFP which is not exported:</p></div></td></tr>
</table>Tree1220http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=391761&oldid=prevTree1220 at 02:49, 18 October 20142014-10-18T02:49:42Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Before we check the functionality of the entire construct, we need to test the YebF protein for its ability to transport attached parts out of the membrane. We began by constructing two circuits:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Before we check the functionality of the entire construct, we need to test the YebF protein for its ability to transport attached parts out of the membrane. We began by constructing two circuits:</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/d/d3/NYMU14_target_YebF_t.png' style='width: 550px;'></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/d/d3/NYMU14_target_YebF_t.png' style='width: 550px;'></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>These circuits are then transformed into <i>E. coli</i>, cultivated in liquid culture, and centrifuged. Afterwards, the supernatants are analyzed for their OD value. If YebF works as we expected, we should see the first circuit successfully secrete RFP into the supernatant :</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>These circuits are then transformed into <i>E. coli</i>, cultivated in liquid culture, and centrifuged. Afterwards, the supernatants are analyzed for <ins class="diffchange diffchange-inline">flourescence intensity and </ins>their OD value. If YebF works as we expected, we should see the first circuit successfully secrete RFP into the supernatant :</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/a/a6/NYMU14_target_YebF_rfp.png' style='width: 220px;margin-left: 40px;'></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/a/a6/NYMU14_target_YebF_rfp.png' style='width: 220px;margin-left: 40px;'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>While the control should only contain RFP which is not exported:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>While the control should only contain RFP which is not exported:</p></div></td></tr>
</table>Tree1220http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=355830&oldid=prevNymuchang at 21:25, 17 October 20142014-10-17T21:25:24Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>According to paper research, we prepared our sample through the protocol described below:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>According to paper research, we prepared our sample through the protocol described below:</p></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li>S.mutans liquid cultured till its stationary phase</li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li>S. mutans liquid cultured till its stationary phase</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Add lysozyme to 5ml cultures</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Add lysozyme to 5ml cultures</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Sonication</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Sonication</li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In this last stage we put all the constructed circuits intended for our probiotic and <i>M102</i> into <i>E. coli</i> and <i>S. mutans</i>, respectively. Then we co-cultured these two transformed strains together to test our full circuit.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In this last stage we put all the constructed circuits intended for our probiotic and <i>M102</i> into <i>E. coli</i> and <i>S. mutans</i>, respectively. Then we co-cultured these two transformed strains together to test our full circuit.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <h1 id='1c-4'>Our <del class="diffchange diffchange-inline">result</del></h1></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <h1 id='1c-4'>Our <ins class="diffchange diffchange-inline">results</ins></h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>nlmC promoter</h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>nlmC promoter</h3></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td></tr>
</table>Nymuchanghttp://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=350208&oldid=prevHiwang123 at 20:24, 17 October 20142014-10-17T20:24:11Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <h>Control-<<del class="diffchange diffchange-inline">font color</del>=<del class="diffchange diffchange-inline">red</del>>Target</<del class="diffchange diffchange-inline">font</del>></h></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <h>Control-<<ins class="diffchange diffchange-inline">span style</ins>=<ins class="diffchange diffchange-inline">'color:rgb(249, 103, 236);'</ins>>Target</<ins class="diffchange diffchange-inline">span</ins>></h></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1>Here’s the Gist…</h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1>Here’s the Gist…</h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class='abstract'></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class='abstract'></div></td></tr>
</table>Hiwang123http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=349828&oldid=prevHiwang123 at 20:20, 17 October 20142014-10-17T20:20:11Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Study done by Liu et al. shown that nlmC promoter can be induced by CSP[1]</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Study done by Liu et al. shown that nlmC promoter can be induced by CSP[1]</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><img src='/wiki/images/d/d9/NYMU14_target_paperres.JPG' style='width: 500px;'></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><img src='/wiki/images/d/d9/NYMU14_target_paperres.JPG' style='width: 500px<ins class="diffchange diffchange-inline">;float: left;padding-right: 10px</ins>;'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>First of all, “constitutive promoter+ RBS+ GFP” and “nlmC promoter+ RBS+ GFP” will be tested by observing the green fluorescence in the gene-modified Streptococcus mutans. “Constitutive promoter+ RBS+ GFP” is believed to create green fluorescence constitutively without addition stimulation. On the other hand, “nlmC promoter+ RBS +GFP” is believed to create green fluorescence under the stimulation of CSP. In this step of function test, we are hoping to test the function of nlmC promoter.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>First of all, “constitutive promoter+ RBS+ GFP” and “nlmC promoter+ RBS+ GFP” will be tested by observing the green fluorescence in the gene-modified Streptococcus mutans. “Constitutive promoter+ RBS+ GFP” is believed to create green fluorescence constitutively without addition stimulation. On the other hand, “nlmC promoter+ RBS +GFP” is believed to create green fluorescence under the stimulation of CSP. In this step of function test, we are hoping to test the function of nlmC promoter.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Secondly, “nlmC promoter+ RBS+ GFP”, “nlmC promoter+ RBS +B0012+ GFP”, and “nlmC promoter +RBS+ B0012+ B0012+GFP” will be tested to see which create the most suitable threshold for preventing cavities. It is believed that the more terminators we added to our circuit, the higher threshold will be made. Under the addition of CSP, the nlmC promoter will be induced, thereby create the green fluorescence which we can observe in the gene-modified Streptococcus mutans.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Secondly, “nlmC promoter+ RBS+ GFP”, “nlmC promoter+ RBS +B0012+ GFP”, and “nlmC promoter +RBS+ B0012+ B0012+GFP” will be tested to see which create the most suitable threshold for preventing cavities. It is believed that the more terminators we added to our circuit, the higher threshold will be made. Under the addition of CSP, the nlmC promoter will be induced, thereby create the green fluorescence which we can observe in the gene-modified Streptococcus mutans.</p></div></td></tr>
</table>Hiwang123http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=349710&oldid=prevHiwang123 at 20:18, 17 October 20142014-10-17T20:18:54Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>Testing Endolysin</h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>Testing Endolysin</h3></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>By doing disc plate assay of endolysin together with several other controls, we can see that there is a clear space besides the disc of our different concentration of our endolysin supernatant.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>By doing disc plate assay of endolysin together with several other controls, we can see that there is a clear space besides the disc of our different concentration of our endolysin supernatant.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p>Here are our samples:</p></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/c/cd/NYMU14_target_endolysin_res1.png' style='width: 370px;float: left;padding: 10px;'></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src='/wiki/images/c/cd/NYMU14_target_endolysin_res1.png' style='width: 370px;float: left;padding: 10px;'></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><p>Here are our samples:</p></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ampicillin : 100ng/ul</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ampicillin : 100ng/ul</li></div></td></tr>
</table>Hiwang123http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=349520&oldid=prevHiwang123 at 20:16, 17 October 20142014-10-17T20:16:51Z<p></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 20:16, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Study done by Liu et al. shown that nlmC promoter can be induced by CSP[1]</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Study done by Liu et al. shown that nlmC promoter can be induced by CSP[1]</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><img src=''></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><img src='<ins class="diffchange diffchange-inline">/wiki/images/d/d9/NYMU14_target_paperres.JPG' style='width: 500px;</ins>'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>First of all, “constitutive promoter+ RBS+ GFP” and “nlmC promoter+ RBS+ GFP” will be tested by observing the green fluorescence in the gene-modified Streptococcus mutans. “Constitutive promoter+ RBS+ GFP” is believed to create green fluorescence constitutively without addition stimulation. On the other hand, “nlmC promoter+ RBS +GFP” is believed to create green fluorescence under the stimulation of CSP. In this step of function test, we are hoping to test the function of nlmC promoter.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>First of all, “constitutive promoter+ RBS+ GFP” and “nlmC promoter+ RBS+ GFP” will be tested by observing the green fluorescence in the gene-modified Streptococcus mutans. “Constitutive promoter+ RBS+ GFP” is believed to create green fluorescence constitutively without addition stimulation. On the other hand, “nlmC promoter+ RBS +GFP” is believed to create green fluorescence under the stimulation of CSP. In this step of function test, we are hoping to test the function of nlmC promoter.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Secondly, “nlmC promoter+ RBS+ GFP”, “nlmC promoter+ RBS +B0012+ GFP”, and “nlmC promoter +RBS+ B0012+ B0012+GFP” will be tested to see which create the most suitable threshold for preventing cavities. It is believed that the more terminators we added to our circuit, the higher threshold will be made. Under the addition of CSP, the nlmC promoter will be induced, thereby create the green fluorescence which we can observe in the gene-modified Streptococcus mutans.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Secondly, “nlmC promoter+ RBS+ GFP”, “nlmC promoter+ RBS +B0012+ GFP”, and “nlmC promoter +RBS+ B0012+ B0012+GFP” will be tested to see which create the most suitable threshold for preventing cavities. It is believed that the more terminators we added to our circuit, the higher threshold will be made. Under the addition of CSP, the nlmC promoter will be induced, thereby create the green fluorescence which we can observe in the gene-modified Streptococcus mutans.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><h3>Testing YebF<h3></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><h3>Testing YebF<<ins class="diffchange diffchange-inline">/</ins>h3></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>From left to right: J23100+yebF+RFP+B0015, J23100, Competent cell</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>From left to right: J23100+yebF+RFP+B0015, J23100, Competent cell</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><img src=''></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><img src='<ins class="diffchange diffchange-inline">/wiki/images/a/a8/NYMU14_target_yefb1.JPG' style='width: 550px;</ins>'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Partially enlargement:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Partially enlargement:</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><img src=''></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><img src='<ins class="diffchange diffchange-inline">/wiki/images/a/a1/NYMU14_target_yefb2.JPG' style='width: 550px;</ins>'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We can see that although there is small difference, there are mostly more fluorescence in our circuit, showing that there might be little RFP secreted. But the effect is not very clear, resulting in small difference in fluorescent intensity.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>We can see that although there is small difference, there are mostly more fluorescence in our circuit, showing that there might be little RFP secreted. But the effect is not very clear, resulting in small difference in fluorescent intensity.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Also, the comparison between our testing circuit with other control shows that there might exist some technical problems, for sometimes LB or competent cell result in more intensity in RFP wavelength range.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Also, the comparison between our testing circuit with other control shows that there might exist some technical problems, for sometimes LB or competent cell result in more intensity in RFP wavelength range.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>At the same time, measuring OD600 to make comparison : </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>At the same time, measuring OD600 to make comparison : </p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><img src=''></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><img src='<ins class="diffchange diffchange-inline">/wiki/images/5/5a/NYMU14_target_yefb3.JPG' style='width: 550px;</ins>'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Overall our testing circuit showed the relationship between growth and its secretion as shown below:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Overall our testing circuit showed the relationship between growth and its secretion as shown below:</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><img src=''></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><img src='<ins class="diffchange diffchange-inline">/wiki/images/f/fc/NYMU14_target_yefb4.JPG' style='width: 550px;</ins>'></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>As seen in the graph, the secretion of YebF and RFP basically soared with the growth of the E.coli, and come to a dynamic balance at the as it entered log phase.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>As seen in the graph, the secretion of YebF and RFP basically soared with the growth of the E.coli, and come to a dynamic balance at the as it entered log phase.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>By doing disc plate assay of endolysin together with several other controls, we can see that there is a clear space besides the disc of our different concentration of our endolysin supernatant.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>By doing disc plate assay of endolysin together with several other controls, we can see that there is a clear space besides the disc of our different concentration of our endolysin supernatant.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Here are our samples:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Here are our samples:</p></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><img src='/wiki/images/c/cd/NYMU14_target_endolysin_res1.png' style='width: 370px;float: left;padding: 10px;'></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ampicillin : 100ng/ul</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ampicillin : 100ng/ul</li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>J23100+yebF+endolysin+B0015 (OD600=2.900)(2.0 rcf, 7min)</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>J23100+yebF+endolysin+B0015 (OD600=2.900)(2.0 rcf, 7min)</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><img src=''></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Western blot filter paper:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Western blot filter paper:</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>AHL induced <i>S. mutans</i> death</h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>AHL induced <i>S. mutans</i> death</h3></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><img src='/wiki/images/c/cb/NYMU14_target_ahl.JPG' style='width: 150px;'></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The supernatant purified from the <i>S. mutans</i> (with constructed circuit: S-IT/PVA838) contains AHL that could possible trigger the killer circuit in <i>E. coli </i>and show a decreasing growth of wild type <i>S. mutans.</i></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The supernatant purified from the <i>S. mutans</i> (with constructed circuit: S-IT/PVA838) contains AHL that could possible trigger the killer circuit in <i>E. coli </i>and show a decreasing growth of wild type <i>S. mutans.</i></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Our constructed circuit (Termed E-C1) including <i>E. coli</i> promoter (Truncated BBa_K523013) region, luxR(BBa_C0062) coding sequence and pLuxR(BBa_R0062) promoter region would be 3117 bp in length. The result of constructed circuit ligation is shown below.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Our constructed circuit (Termed E-C1) including <i>E. coli</i> promoter (Truncated BBa_K523013) region, luxR(BBa_C0062) coding sequence and pLuxR(BBa_R0062) promoter region would be 3117 bp in length. The result of constructed circuit ligation is shown below.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><img src=''></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></div> <!-- article --> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></div> <!-- article --> </div></td></tr>
</table>Hiwang123http://2014.igem.org/wiki/index.php?title=Team:NYMU-Taipei/project/1c&diff=347128&oldid=prevHiwang123 at 19:52, 17 October 20142014-10-17T19:52:37Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 19:52, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1 id='1c-4'>Our result</h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1 id='1c-4'>Our result</h1></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><h3>nlmC promoter</h3></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>Constitutive promoter+ RBS+ GFP</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>nlmC promoter+ RBS+ GFP</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>nlmC promoter+ RBS+B0012+GFP</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>nlmC promoter+ RBS+ B0012+B0012+GFP</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Study done by Liu et al. shown that nlmC promoter can be induced by CSP[1]</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>First of all, “constitutive promoter+ RBS+ GFP” and “nlmC promoter+ RBS+ GFP” will be tested by observing the green fluorescence in the gene-modified Streptococcus mutans. “Constitutive promoter+ RBS+ GFP” is believed to create green fluorescence constitutively without addition stimulation. On the other hand, “nlmC promoter+ RBS +GFP” is believed to create green fluorescence under the stimulation of CSP. In this step of function test, we are hoping to test the function of nlmC promoter.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Secondly, “nlmC promoter+ RBS+ GFP”, “nlmC promoter+ RBS +B0012+ GFP”, and “nlmC promoter +RBS+ B0012+ B0012+GFP” will be tested to see which create the most suitable threshold for preventing cavities. It is believed that the more terminators we added to our circuit, the higher threshold will be made. Under the addition of CSP, the nlmC promoter will be induced, thereby create the green fluorescence which we can observe in the gene-modified Streptococcus mutans.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><h3>Testing YebF<h3></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>From left to right: J23100+yebF+RFP+B0015, J23100, Competent cell</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Partially enlargement:</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>We can see that although there is small difference, there are mostly more fluorescence in our circuit, showing that there might be little RFP secreted. But the effect is not very clear, resulting in small difference in fluorescent intensity.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Also, the comparison between our testing circuit with other control shows that there might exist some technical problems, for sometimes LB or competent cell result in more intensity in RFP wavelength range.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>At the same time, measuring OD600 to make comparison : </p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Overall our testing circuit showed the relationship between growth and its secretion as shown below:</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>As seen in the graph, the secretion of YebF and RFP basically soared with the growth of the E.coli, and come to a dynamic balance at the as it entered log phase.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><h3>Testing Endolysin</h3></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>By doing disc plate assay of endolysin together with several other controls, we can see that there is a clear space besides the disc of our different concentration of our endolysin supernatant.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Here are our samples:</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>Ampicillin : 100ng/ul</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>BHI broth</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>J23100+yebF+B0015: J23100+yebF+B0015 overnight culture supernatant</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(OD600=2.647)(2.0 rcf, 7min) </li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>J23100+yebF+endolysin+B0015:overnight culture supernatant </li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>J23100+yebF+endolysin+B0015 (OD600=2.900)(2.0 rcf, 7min)</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Western blot filter paper:</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>Positive control: Ampicillin (100ng/ul),50ml</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>Negative control: BHI, 50ml /J23100+yebF+B0015, 50ml</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>Testing:</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li>J23100+yebF+endolysin+B0015, 50ml</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li>J23100+yebF+endolysin+B0015, 100ml</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>As shown in the figure, ampicillin showed a significant inhibition zone as a positive control. Three positive control all show there ability to kill <i>Streptococcus mutans</i> by the clear zone around the disc. The BHI negative control showed no killing ability as expected.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>However, there are also little <i>Streptococcus mutans</i> growing near J23100+yebF+B0015 culture supernatant. By observing the pattern on the plate, we speculated that it might be the result of the diffusion of Ampicillin beside it.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><h3>AHL induced <i>S. mutans</i> death</h3></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>The supernatant purified from the <i>S. mutans</i> (with constructed circuit: S-IT/PVA838) contains AHL that could possible trigger the killer circuit in <i>E. coli </i>and show a decreasing growth of wild type <i>S. mutans.</i></p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Our constructed circuit (Termed E-C1) including <i>E. coli</i> promoter (Truncated BBa_K523013) region, luxR(BBa_C0062) coding sequence and pLuxR(BBa_R0062) promoter region would be 3117 bp in length. The result of constructed circuit ligation is shown below.</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><img src=''></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></div> <!-- article --> </ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1 id='1c-5'>Reference</h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1 id='1c-5'>Reference</h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ol></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ol></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li><del class="diffchange diffchange-inline">http</del>:<del class="diffchange diffchange-inline">//www</del>.<del class="diffchange diffchange-inline">expertsmind</del>.<del class="diffchange diffchange-inline">com</del>/<del class="diffchange diffchange-inline">topic</del>/<del class="diffchange diffchange-inline">microbiology</del>/<del class="diffchange diffchange-inline">bacterial-cell-wall-92313.aspx</del></li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li><ins class="diffchange diffchange-inline">Liu, T., et al., ComCED signal loop precisely regulates nlmC expression in Streptococcus mutans. Annals of Microbiology, 2014. 64(1)</ins>: <ins class="diffchange diffchange-inline">p</ins>. <ins class="diffchange diffchange-inline">31-38</ins>.<ins class="diffchange diffchange-inline"><</ins>/<ins class="diffchange diffchange-inline">li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(2006)"Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli"<</ins>/<ins class="diffchange diffchange-inline">li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(2007)"ACES™ Signal Sequence and YebF Expression Systems Technical Brief"<</ins>/<ins class="diffchange diffchange-inline">li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(2012)"A Protein Export Pathway Involving Escherichia coli Porins"</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(2006)patent" PROTEIN PRODUCTION METHOD UTILIZING YEBF "</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(2007)"Genome sequence of Streptococcus mutans bacteriophage M102"</li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li>(2012)''Biology and Genome Sequence of Streptococcus mutans Phage M102AD''</ins></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ol></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ol></div></td></tr>
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